A NOVEL METHOD FOR THE AMINO-ACID-SEQUENCE CONFIGURATION DETERMINATION OF PEPTIDES CONTAINING D/L-AMINO ACIDS UTILIZING A FLUOROGENIC EDMAN REAGENT, 7-N,N-DIMETHYLAMINOSULPHONYL-4-(2,1,3-BENZOXADIAZOLYL)ISOTHIOCYANATE (DBD-NCS)

被引:18
作者
IMAI, K [1 ]
MATSUNAGA, H [1 ]
FUKUSHIMA, T [1 ]
SANTA, T [1 ]
HOMMA, H [1 ]
NAKASHIMA, K [1 ]
AKIYAMA, S [1 ]
机构
[1] NAGASAKI UNIV,SCH PHARMACEUT SCI,NAGASAKI 852,JAPAN
关键词
D O I
10.1002/bmc.1130090309
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A novel method for the amino acid sequence and configuration determination of peptides containing D- or L-amino acids is presented. An enkephalin analogue, [D-Ala(2), D-Leu(5)]enkephalin (Tyr-Ala-Gly-Phe-Leu) was derivatized with a fluorogenic Edman reagent, 7-N,N-dimethylaminosulphonyl-4-(2,1,3-benzoxadiazolyl) isothiocyanate (DBD-NCS), cleaved and cyclized with trifluoroacetic acid at 50 degrees C for 1 min and the resultant thiazolinone derivative (DBD-thiazolinyl-L-Tyr) was separated from the racemized DBD-thiazolinyl-D-Tyr (about 20% as compared to the L-isomer) on a phenylcarbamylated cyclodextrin column: The separation factor (alpha) for D- and L-isomers was 1.09. The column eluate was monitored fluorometrically at 524 nm with excitation at 387 nm. The detection limit was about pmol range. The same treatment adopted for the residual peptide, Ala-Gly-Phe-Leu, gave DBD-thiazolinyl-D-Ala (alpha = 1.09 for D,L-Phe) with the lesser amount of L-analogue (about 20%). In the same manner, Gly and L-Phe (alpha = 1.09) were detected. The method might be useful for the study of aging of proteins such as eye lens and the amyloid proteins derived from Alzheimer's disease.
引用
收藏
页码:152 / 154
页数:3
相关论文
共 9 条
  • [1] CHIRAL ANALYSIS OF THE REACTION STAGES IN THE EDMAN METHOD FOR SEQUENCING PEPTIDES
    DAVIES, JS
    MOHAMMED, AKA
    [J]. JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 2, 1984, (10): : 1723 - 1727
  • [2] PREPARATION OF PHENYL THIOHYDANTOINS FROM SOME NATURAL AMINO ACIDS
    EDMAN, P
    [J]. ACTA CHEMICA SCANDINAVICA, 1950, 4 (02): : 277 - 282
  • [3] RACEMIZED D-ASPARTATE IN ALZHEIMER NEUROFIBRILLARY TANGLES
    FISHER, GH
    PAYAN, IL
    CHOU, SJ
    MAN, EH
    CERWINSKI, S
    MARTIN, T
    EMORY, C
    FREY, WH
    [J]. BRAIN RESEARCH BULLETIN, 1992, 28 (01) : 127 - 131
  • [4] D-ASPARTIC ACID IN PURIFIED MYELIN AND MYELIN BASIC-PROTEIN
    FISHER, GH
    GARCIA, NM
    PAYAN, IL
    CADILLAPEREZRIOS, R
    SHEREMATA, WA
    MAN, EH
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1986, 135 (02) : 683 - 687
  • [5] FUKUSHIMA K, 1994, IN PRESS ANALYST
  • [6] SYNTHESIS OF NOVEL FLUOROGENIC EDMAN REAGENTS, 7-N,N-DIMETHYLAMINOSULPHONYL-4-(2,1,3-BENZOXADIAZOLYL)ISOTHIOCYANATE (DBD-NCS) AND 7-AMINOSULPHONYL-4-(2,1,3-BENZOXADIAZOLYL)-ISOTHIOCYANATE (ABD-NCS)
    IMAI, K
    UZU, S
    NAKASHIMA, K
    AKIYAMA, S
    [J]. BIOMEDICAL CHROMATOGRAPHY, 1993, 7 (01) : 56 - 57
  • [7] ASPARTIC-ACID RACEMIZATION IN HUMAN LENS DURING AGING AND IN CATARACT FORMATION
    MASTERS, PM
    BADA, JL
    ZIGLER, JS
    [J]. NATURE, 1977, 268 (5615) : 71 - 73
  • [8] D-ALANINE IN THE FROG-SKIN PEPTIDE DERMORPHIN IS DERIVED FROM L-ALANINE IN THE PRECURSOR
    RICHTER, K
    EGGER, R
    KREIL, G
    [J]. SCIENCE, 1987, 238 (4824) : 200 - 202
  • [9] DETERMINATION OF THE CHIRALITY OF AMINO-ACID-RESIDUES IN THE COURSE OF SUBTRACTIVE EDMAN DEGRADATION OF PEPTIDES
    SCALONI, A
    SIMMACO, M
    BOSSA, F
    [J]. ANALYTICAL BIOCHEMISTRY, 1991, 197 (02) : 305 - 310